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Issue 10, 2013
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Selective labeling of mature RISC using a siRNA carrying fluorophore–quencher pair

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Abstract

RNA interference (RNAi) is an endogenous gene silencing system that has been harnessed to inhibit expression of specific genes through the introduction of short double-stranded RNAs, called small interfering RNAs (siRNAs) into cells. After entry into a cell, an siRNA is assembled into the RNA-induced silencing complex (RISC) and suppresses the target gene translation through interaction between the antisense (guide) strand of the siRNA and the target mRNA. To evaluate the intracellular fate of siRNAs, we performed imaging analyses using siRNAs labeled with newly designed fluorophore–quencher clusters introduced through the base surrogate with D-threoninol as a scaffold. Fluorescence microscopy analyses indicated that mature RISC containing fluorescently labeled antisense strand was localized within P-bodies. Thus, the antisense strand uptake into the RISC machinery was successfully visualized.

Graphical abstract: Selective labeling of mature RISC using a siRNA carrying fluorophore–quencher pair

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Supplementary files

Article information


Submitted
02 May 2013
Accepted
25 Jul 2013
First published
26 Jul 2013

Chem. Sci., 2013,4, 4016-4021
Article type
Edge Article

Selective labeling of mature RISC using a siRNA carrying fluorophore–quencher pair

Y. Kamiya, A. Ito, H. Ito, M. Urushihara, J. Takai, T. Fujii, X. Liang, H. Kashida and H. Asanuma, Chem. Sci., 2013, 4, 4016
DOI: 10.1039/C3SC51197A

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